5244results about "DNA preparation" patented technology

The invention provides an UTR (Untranslated Region) element H2202 P1-G as well as a construction method and application thereof, and relates to the technical field of mRNA (messenger ribonucleic acid). According to the present invention, the ribosome load prediction and the secondary structure optimization are performed on the natural 5 'UTR of the HIV TAT 202 gene through the BaidleHelix platform, and the obtained HTAT 202 P1 sequence avoids the inhibitory hairpin structure so as to significantly improve the luciferase expression quantity compared to the natural UTR; an ncRNA sequence without a secondary structure is introduced on the basis of the HTAT 202 P1, translation inhibition of a 5 'cap region is further relieved, and the protein expression quantity of the constructed H2202 P1-G mutant (the DNA sequence of the H2202 P1-G is as shown in SEQ NO 1, and the RNA sequence is as shown in SEQ NO 2) is further improved.

A sample holder includes a first member featuring a first retaining mechanism configured to retain a first substrate that includes a sample, a second member featuring a second retaining mechanism configured to retain a second substrate that includes a reagent medium, and an alignment mechanism connected to at least one of the first and second members, and configured to align the first and second members such that the sample contacts at least a portion of the reagent medium when the first and second members are aligned.

The invention provides an UTR element NHP1 as well as a construction method and application thereof, and relates to the technical field of mRNA. A 5 'UTR with a good expression effect is designed by integrating dominant sequences of a human high-expression gene and a pathogen natural UTR, a chimeric structure NHP1 with high ribosome load is predicted through a calculation model, a DNA sequence of the NHP1 is as shown in SEQ NO 1, and an RNA sequence of the NHP1 is as shown in SEQ NO 2; an EGFP report system is adopted on the DNA level to rapidly screen UTR; the translation efficiency is quantitatively evaluated on the RNA level through luciferase mRNA (N1-methyl pseudouridine modification); and the particle size is controlled by a microfluidic technology, so that the optimized UTR-mRNA is efficiently expressed after being delivered.

This disclosure provides methods for spatial profiling of biological analytes present in a biological sample. Methods include generating feature arrays using a master / copy format using recessed arrays, and methods for using such arrays. For example spatially-tagged analyte capture analytes can be used in spatial detection in methods to determine the location of analytes (e.g., proteins) in biological samples.

A sample holder includes a first member featuring a first retaining mechanism configured to retain a first substrate that includes a sample, a second member featuring a second retaining mechanism configured to retain a second substrate that includes a reagent medium, and an alignment mechanism connected to at least one of the first and second members, and configured to align the first and second members such that the sample contacts at least a portion of the reagent medium when the first and second members are aligned.

The invention belongs to the field of prevention and treatment of tilletia controversa Kuhn, and particularly relates to a tilletia controversa Kuhn effect protein g16561 and application thereof. An amino acid sequence of the Tilletia controversa Kuhn effect protein is shown as SEQ ID NO: 2. The effect protein plays an important role in the process of inhibiting plant defense reaction, and has an interaction relationship with a wheat protein in the process of infecting wheat by Tilletia controversa Kuhn. According to the invention, the pathogenesis of Tilletia controversa Kuhn can be deeply researched, and the excavation of wheat disease-resistant genes and the cultivation of wheat disease-resistant varieties can be promoted.

The invention provides a nucleic acid aptamer for specific recognition of morphine and application of the nucleic acid aptamer, and belongs to the technical field of biosensing and detection. The nucleotide sequence of the nucleic acid aptamer is as shown in SEQ ID NO: 1. The screening method is based on a Capture-SELEX technology and comprises the key steps that streptavidin magnetic beads are used for fixing an ssDNA library, estradiol, deabietic acid and totarol are introduced to serve as reverse screening substances so as to remove non-specific sequences, and finally the high-specificity aptamer is obtained through high-throughput sequencing and affinity determination. The dissociation constant of the aptamer and morphine is 127.31 nM, and the aptamer shows high affinity and high specificity. The invention further relates to application of the aptamer in preparation of a sensor and a kit for detecting morphine, and a new technical means is provided for rapid detection of morphine.

Methods for producing a linear deoxyribonucleic acid (DNA) product with enhanced resistance to nuclease digestion are provided. The methods comprise, (a) digesting a double-stranded DNA molecule with an endonuclease that cleaves an endonuclease target sequence to generate a digested double-stranded DNA molecule, wherein the digested double-stranded DNA molecule comprises a linear double-stranded region, and a truncated protelomerase sequence at a first end, wherein the truncated protelomerase target sequence is non-functional; (b) appending a first adaptor molecule to the first end of the digested double-stranded DNA molecule and appending a second adaptor molecule to the second end of the digested double-stranded DNA molecule to generate a precursor double-stranded DNA molecule, wherein the first adaptor molecule comprises a truncated protelomerase target sequence that forms a first functional protelomerase target sequence with the truncated protelomerase sequence at the first end of the digested double-stranded DNA molecule; and (c) incubating the precursor double-stranded DNA molecule with a protelomerase to generate the linear DNA product, wherein the protelomerase closes the first end of the precursor double-stranded DNA molecule at the first functional protelomerase target sequence.

The present invention relates to a nuclease polypeptide, such as a CRISPR nuclease polypeptide derived from a reference nuclease, which may be nuclease A, nuclease K or nuclease M, comprising a RuvC nuclease domain and a HNH nuclease domain. Also provided herein are gene editing systems comprising such nuclease polypeptides and gene editing methods using the gene editing systems.

Methods for determining a location of a feature in a spatial array with features include: (a) providing an array with a first set of one or more features immobilized on a substrate, a first feature of the first set having a first barcoded oligonucleotide with a first spatial barcode and a first constant sequence, and a second set of one or more features immobilized on the substrate, a second feature of the second set having a second barcoded oligonucleotide with a second spatial barcode and a second constant sequence; (b) attaching the first constant sequence to the second constant sequence to generate a nucleic acid product; (c) determining all or a portion of a sequence of the nucleic acid product or a complement thereof; and (d) associating the second barcoded oligonucleotide with the first barcoded oligonucleotide in the nucleic acid product.

The invention discloses a nucleic acid aptamer OUC-Fx11-5T of fucoxanthin and application of the nucleic acid aptamer OUC-Fx11-5T, and belongs to the technical field of nucleic acid aptamers. The nucleic acid aptamer OUC-Fx11-5T of the fucoxanthine has a nucleotide sequence as shown in SEQ ID NO. 19. The nucleic acid aptamer OUC-Fx11-5T of the fucoxanthine has a nucleotide sequence as shown in SEQ The invention further discloses the application of the nucleic acid aptamer OUC-Fx11-5T of the fucoxanthin in identification or detection of the fucoxanthin. According to the nucleic acid aptamer OUC-Fx11-5T of the fucoxanthine, the affinity and dissociation constant of the nucleic acid aptamer OUC-Fx11-5T and the fucoxanthine is determined to be 68.2 nM through an isothermal titer thermal method, the nucleic acid aptamer OUC-Fx11-5T has no obvious affinity to structural analogues of the fucoxanthine such as violaxanthin, neoxanthin, zeaxanthin and beta-carotene, has high affinity and good specificity to the fucoxanthine, and can be used for preparing the fucoxanthine aptamer OUC-Fx11-5T of the fucoxanthine. The method can be used for identification or detection of fucoxanthine.

The invention provides a kit for rapidly detecting MRSA and drug-resistant gene nucleic acid POCT of the MRSA based on a phage lyase mutant. The kit comprises nucleic acid extraction and purification and triple PCR rapid amplification detection reagents. The staphylococcus aureus bacteriophage lyase mutant nucleic acid release liquid containing the amino acid sequence as shown in SEQ ID NO: 1 is adopted, wall breaking can be rapidly conducted on staphylococcus aureus, and nucleic acid is released. The effect of the nucleic acid release liquid acting for 5 minutes is equivalent to or even better than that of the lysozyme acting for 60 minutes; according to the present invention, with the combination of the MRSA and drug-resistant gene triple PCR rapid amplification detection reagent, the total detection time is only 35 min, the time is saved by 55 min or more than 55 min compared with the lysozyme method, and the MRSA and drug-resistant gene nucleic acid POCT rapid detection can be achieved;

The invention discloses a preparation method of DNA (Deoxyribonucleic Acid) hydrogel with programmable mechanical strength. By regulating and controlling the molar ratio of the catalytic initiating chains to the total amount of the DNA monomer building blocks, multiple hydrogels which are equivalent in mass concentration and have different mechanical strengths are constructed. According to the method, a strategy similar to catalytic assembly is adopted, the feeding molar ratio of a catalytic initiating chain to a DNA monomer building block is creatively adjusted, the proportion of cross-linking reactions at different levels is regulated and controlled, then the DNA hydrogel with different mechanical strengths is synthesized under the isothermal condition, and the storage modulus range is about 200-2000 Pa. In addition, the change of the mechanical strength is independent of the total concentration of the matrix (DNA) of the hydrogel, the DNA sequence, the stress relaxation rate and other properties. The method provides a novel convenient method for constructing DNA hydrogels with different mechanical strengths, and possibly provides a novel tool for researching the specific influence of mechanical properties of materials on cell / tissue behaviors.

Methods and compositions for modulating repeat non-ATG protein (RAN protein) translation are provided. In some aspects, the disclosure provides methods of inhibiting RAN protein translation by contacting a cell with an effective amount of an inhibitor of eIF2 phosphorylation or an inhibitor of protein kinase R (PKR). In some embodiments, methods described by the disclosure are useful for treating diseases associated with RAN protein translation, such as certain neurodegenerative diseases.

Disclosed herein is an aqueous nucleic acid purification buffer comprising a polar aprotic solvent. Also provided herein are using such purification buffers to precipitate nucleic acids from a solution onto a solid support, thereby providing a solid support that binds to nucleic acids. Also provided herein is a method for treating a nucleic acid comprising exposing a sample comprising the nucleic acid to an aqueous medium comprising a polar aprotic solvent in the presence of a solid support; and precipitating the nucleic acid onto the solid support, thereby providing a solid support that binds to the nucleic acid. Further provided herein are kits and compositions comprising such aqueous nucleic acid purification buffers, as well as related nucleic acid analysis devices, and uses of the kits.

The present disclosure provides systems and methods for separating and characterizing oligonucleotides. Fixed dose combinations of small interfering RNAs can be subjected to denaturing ion pairing reverse phase liquid chromatography using optimal conditions to separate single RNA strands for purity analysis.

The invention belongs to the technical field of gene engineering, and discloses a CRISPR / Cas9 delivery system and application thereof. The CRISPR / Cas9 delivery system is obtained by loading the CRISPR / Cas9 system with the fluorinated polyethyleneimine, and the fluorinated polyethyleneimine can deliver the CRISPR / Cas9 carrier into cells, has excellent transmembrane delivery efficiency, and can efficiently mediate the CRISPR / Cas9 system to enter target cells. The CRISPR / Cas9 delivery system is applied to large-scale breeding of aquatic animals, the use safety is high, the biocompatibility is excellent, and the delivery system has excellent gene editing efficiency and has wide application prospects in the field of gene editing breeding.

The invention provides a specific sequence nucleic acid molecule and application thereof as a PDRN, and relates to the technical field of biology. A novel nucleic acid molecule with a specific sequence is obtained, and the sequence has certainty, biological activity and stable product quality. The PDRN of the specific sequence does not have cytotoxicity, is high in safety and has a wide application prospect. Moreover, compared with the extracted PDRN, the PDRN with a specific sequence is found for the first time, the product has a remarkable effect of promoting VEGF expression, cell damage can be effectively repaired, and a novel skin-care and repairing functional component is provided for the industries of cosmetics and medicines. The invention further provides a preparation method of the PDRN with the specific sequence, and the preparation method is simple and suitable for popularization, application and industrial production.

Methods for determining a location of a feature on an array include: (a) providing a first array with a first plurality of features immobilized on a first substrate; (b) providing a second array with a second plurality of features immobilized on a second substrate; (c) aligning the first array with the second array; (d) hybridizing a first barcoded oligonucleotide of the first array to a second barcoded oligonucleotide of the second array, thereby producing a combined nucleic acid that includes first and second spatial barcodes; (e) determining all or a portion of the sequence of the combined nucleic acid; and (f) identifying the second barcoded oligonucleotide associated with the first barcoded oligonucleotide in the combined nucleic acid, and determining the location of a second feature in the second array.

The invention relates to the field of gene engineering. In particular, the present invention relates to cytosine deaminases and their use in base editing. More specifically, the invention relates to a method for screening and identifying a deaminase, a base editing system based on a newly identified cytosine deaminase, a method for editing a target sequence in a genome of an organism (such as a plant) by using the base editing system, and a method for screening and identifying the target sequence. As well as genetically modified organisms (e.g., plants) and progeny thereof produced by the method.

The invention provides a nucleic acid aptamer and a screening method thereof, the aptamer is of an oligonucleotide DNA structure, the nucleotide sequence of the nucleic acid aptamer is the nucleotide sequence of any DNA fragment as shown in SEQ ID NO: 1-9, and the nucleic acid aptamer can specifically target cardiac muscle cells (CMs). According to the invention, a new strategy can be provided for clinical treatment of cardiovascular diseases, and meanwhile, bioactive molecules such as microRNA (miRNA), small interfering RNA (small interfering RNA, siRNA), lipidosome, microspheres, microcapsules and the like can be carried to be used as an intracellular drug delivery tool.

Disclosed herein are predictive models for T-cell epitope prediction, B-cell epitope prediction, and protein design wherein a method is implemented for generating a protein variant amino acid sequence of a target protein having one or more modified properties, the method comprising: (a) iteratively sampling an input amino acid sequence of the target protein, and (b) sampling the individual protein score of at least one weighted relative contribution of the single residue mutant input amino acid sequence to the at least one target property across a plurality of other single residue mutant input amino acid sequences to generate a combined protein score, wherein the combined protein score corresponds to the protein variant comprising one or more amino acid mutations of the single residue mutant input amino acid sequences.

The invention relates to a nucleic acid aptamer for specifically recognizing tilmicosin as well as screening and application of the nucleic acid aptamer, and belongs to the technical field of food safety detection and biosensing. Aiming at the defects of the existing method for detecting tilmicosin residues in food, the invention provides a high-affinity and high-specificity nucleic acid aptamer obtained by screening based on a graphene oxide index enrichment ligand systematic evolution technology. The aptamer is obtained by multiple rounds of screening from a random single-stranded DNA library through a GO-SELEX technology, and a core sequence Apt-2-1 with the length of 33 nt is finally obtained through sequence analysis and structure optimization. The aptamer shows excellent binding capacity to tilmicosin, the dissociation constant reaches 9.82 nM, and the aptamer has good specificity. The nucleic acid aptamer provided by the invention can be used for constructing a rapid and sensitive tilmicosin residue detection method, and has important application value in monitoring of livestock and poultry products and food safety.

Protected DNA comprising a single-stranded DNA (ssDNA) cassette is provided. Further provided are uses of the protected DNA, methods for producing protected DNA, products generated in performing such methods (including intermediate and final products), and kits for use in such methods.

An automated high throughput screening platform for identification of compounds and genomic constructs having mitochondrial function modulating activity and compounds so identified are provided. Use of such compounds for modulating mitochondrial function are also disclosed. A high-throughput screening method of determining genetic mechanisms of action for therapeutic or toxic agents in genetic or pharmacologic models of disease are also provided.

Methods for determining a location of a feature on a spatial array include (a) providing an array of features on a substrate, where a feature of the array includes a barcoded oligonucleotide having, in a 5′ to 3′ direction, a spatial barcode, a cleavage domain, and a constant sequence; (b) hybridizing a priming oligonucleotide to the constant sequence; (c) extending the priming oligonucleotide using the barcoded oligonucleotide as a template; and (d) determining all or a portion of a sequence of the extended priming oligonucleotide corresponding to the spatial barcode, or a complement thereof, and a location of the extended priming oligonucleotide, and using the location of the extended priming oligonucleotide to determine the location of the feature on the spatial array.

The present disclosure relates to Taq DNA Polymerase mutants comprising at least three mutations which are capable of high levels of DNA amplification in the presence of several PCR inhibitors and compositions comprising the same. Also disclosed are methods of performing mutagenesis on wild-type Taq DNA polymerase to obtain the disclosed Taq DNA polymerase mutants, DNA amplification using the same, and cDNA amplification using the same.

The present disclosure relates to methods of identifying RNA targets of RNA binding proteins. In aspects, the disclosure relates to a method of identifying RNA molecules bound by RNA binding proteins. Some embodiments of the present disclosure relate to a method that can definitively identify direct RNA-target interactions with targeted proteins without the requirement for immunoprecipitation or gel extraction. In some embodiments, the method may include combining multiple antibodies in the same sample.

Compositions and methods for editing, e.g., introducing double-stranded breaks, within the KLKB1 gene are provided. Compositions and methods for treating subjects having hereditary angioedema (HAE), are provided.